bFGF对3-硝基丙酸处理的小鼠受精卵体外发育能力的影响  

Effect of bFGF on in vitro Development of Mouse ZygotesTreated with 3-nitropropionic Acid

作　　者：田微 方妍雅 柳海星[1] 李钟淑[1] TIAN Wei;FANG Yanya;LIU Haixing;LI Zhongshu(Agricultural College of Yanbian University,Yanji 133000,China)

机构地区：[1]延边大学农学院,延吉133000

出　　处：《中国畜牧兽医》2022年第2期608-614,共7页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金“氧化应激诱导的母型-合子型时期哺乳动物胚胎发育阻滞机理的研究”(32060756);国家自然科学基金“哺乳动物体细胞克隆胚胎抗氧化机理的研究”(31360546);延边州延边黄牛品种选育项目(413090128、413090132、413090127)。

摘　　要：【目的】研究在体外培养液中添加碱性成纤维细胞生长因子(bFGF)对3-硝基丙酸(3-NPA)处理的小鼠受精卵体外发育能力的影响,为提高氧化应激早期胚胎体外发育质量提供参考。【方法】在小鼠受精卵体外培养液中添加0、20、50、100和150 ng/mL bFGF,培养24、48和96 h,统计2-细胞率、4-细胞率和囊胚率,筛选最佳bFGF处理浓度。经腹腔注射12.5 mg/kg 3-NPA生产氧化应激体内受精卵,正常组小鼠腹腔注射等体积生理盐水,将获得的受精卵分为添加或不添加bFGF组,即3-NPA和3-NPA+bFGF组及对照组(C)和bFGF组,培养到囊胚后,用DCFH-DA检测胚胎内活性氧(reactive oxygen species,ROS)水平,CMF2HC检测谷胱甘肽(glutathione,GSH)水平,JC-1检测早期胚胎线粒体膜电位强度。【结果】0、20、50、100和150 ng/mL bFGF组2-细胞率均无显著差异(P>0.05),100 ng/mL bFGF组囊胚率显著高于其他各组(P<0.05),150 ng/mL bFGF组4-细胞率和囊胚率均显著低于其他各组(P<0.05),因此后续试验选用100 ng/mL bFGF。3-NPA+bFGF组4-细胞率显著高于3-NPA组(P<0.05),bFGF组囊胚率显著高于其他组(P<0.05)。bFGF+3-NPA组囊胚率显著高于3-NPA组(P<0.05);与对照组相比,3-NPA组ROS水平显著升高(P<0.05),bFGF组ROS水平显著降低(P<0.05);3-NPA组GSH和线粒体膜电位水平均显著降低(P<0.05),bFGF组GSH和线粒体膜电位水平均显著增加(P<0.05)。bFGF+3-NPA组ROS水平显著低于3-NPA组(P<0.05),GSH和线粒体膜电位水平均显著高于3-NPA组(P<0.05)。【结论】在体外培养液中添加100 ng/mL bFGF可减少3-NPA诱导的胚胎氧化应激、改善胚胎线粒体功能,从而提高小鼠受精卵体外发育的能力。【Objective】This experiment was to study the effect of adding basic fibroblast growth factor(bFGF)to the in vitro development medium of mouse fertilized eggs treated with 3-nitropropionic acid(3-NPA),in order to provide reference for improving the in vitro development quality of early embryos under oxidative stress.【Method】0,20,50,100 and 150 ng/mL bFGF were added to the in vitro culture medium of mouse zygotes,cultured for 24,48 and 96 h,and the 2-cell rate,4-cell rate and blastocyst rate were counted to screen the best bFGF treatment concentration.In vivo zygotes were produced by intraperitoneal injection of 12.5 mg/kg 3-NPA,and the same volume of normal saline was injected intraperitoneally as the control,the fertilized eggs were divided into 3-NPA and 3-NPA+bFGF groups,control group(C)and bFGF group.After blastocysts were obtained,the level of reactive oxygen species(ROS)in embryos was detected by DCFH-DA,the level of glutathione(GSH)was detected by CMF2HC,and the mitochondrial membrane potential intensity of early embryos was detected by JC-1.【Results】There was no significant difference in 2-cell rate among 0,20,50,100 and 150 ng/mL bFGF groups(P>0.05),and the blastocyst rate in 100 ng/mL bFGF group was significantly higher than that of the other groups(P<0.05),the 4-cell rate and blastocyst rate in 150 ng/mL bFGF group were significantly lower than those in the other groups(P<0.05),so 100 ng/mL bFGF was used in subsequent experiments.The 4-cell rate in 3-NPA+bFGF group was significantly higher than that in 3-NPA group(P<0.05),the blastocyst rate in bFGF group was significantly higher than that of the other groups(P<0.05).The blastocyst rate in bFGF+3-NPA group was significantly higher than that in 3-NPA group(P<0.05).Compared with control group,the level of ROS in 3-NPA group was increased significantly(P<0.05),and the level of ROS in bFGF group was decreased significantly(P<0.05).GSH and mitochondrial membrane potential were decreased significantly in 3-NPA group(P<0.05),and were increased signi

关 键 词：小鼠 胚胎发育 碱性成纤维细胞生长因子(bFGF) 3-硝基丙酸(3-NPA) 活性氧 

分 类 号：S813.3[农业科学—畜牧学]